Kaneka Corporation

Kanegafuchi Kagaku Kogyo Kabushiki Kaisha

Kanegafuchi Chemical Industry Co., Ltd.

Yasuhiro Ikenaka

Yukio Yamada

Kazuyoshi Yajima

Satomi Takahashi

Masayuki Takano

Hirokazu Nanba

Junzo Hasegawa

Akira Iwasaki

Noriyuki Kizaki

Masahiro Ogura

Hideyuki Matsuda

Makoto Kawamukai

Sakayu Shimizu

Hideaki Yamada

Kiyoshi Watanabe

Yoshihiko Yasohara

Keiji Matsumoto

Takehisa Ohashi

Hajime Kawaharada

Tetsu Kakutani

Kenji Yamashita

Kazuhiko Yamada

Yoshiro Hiraishi

Kenichi Nishi

Kazuma Okubo

The invention aims at providing a process for producing coenzyme Qefficiently in microorganisms by utilizing a coenzyme Qside chain synthesis gene derived from a fungal species belonging to the genusand genus The present invention relates to a DNA having a DNA sequence described under SEQ ID NO:1 and 2 or derived from the above sequence by deletion, addition, insertion and/or substitution of one or several bases and encoding a protein having decaprenyl diphosphate synthase activity.

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To provide a method for preparing optically active compounds, which mainly contain (R)-amino compounds, by microbial enzymes efficiently and inexpensively; a polypeptide having stereoselective transaminase activity which can be suitably used for the above preparation method; and a DNA encoding the polypeptide. A method for preparing an optically active amino compound, characterized in stereoselectively transaminating by acting a transaminase on an amino group acceptor, a ketone compound in the presence of an amino group donor, a primary amine; a DNA comprising a nucleotide sequence encoding a polypeptide having stereoselective transaminase activity; and a polypeptide having stereoselective transaminase activity obtainable from a culture of a microorganism belonging to the genus Arthrobacter.

Producing optically active amino compounds

Process for producing coenzyme Q10

A gene of decaprenyl diphosphate synthase, which is the key gene participating in the biosynthesis of coenzyme Q was isolated from a bacterium belonging to the family Rhizobiaceae. By transferring this gene into a microorganism such as and expressing therein, coenzyme Q can be effectively produced.

Relating to an enzyme capable of efficiently converting a ketone compound to an optically active amino compound by transamination, and a process for preparing an optically active amino compound using the enzyme. An (S)-&agr;-phenethylamine:pyruvate transaminase, which acts on (S)-&agr;-phenethylamine and a ketone compound, thereby catalyzing transamination for forming acetophenone and an amino compound corresponding to the ketone compound; a process for preparing an optically active amino compound using the transaminase; and a method for culturing a microorganism for producing the above transaminase, comprising adding to a medium one or more compounds selected from the group consisting of propylamine, 1-butylamine, 2-butylamine, 2-pentylamine, isopropylamine and isobutylamine as an inducer for the enzyme when a microorganism for producing (S)-&agr;-phenethylamine:pyruvate transaminase is cultured.

(S)-&agr;-phenethylamine: pyruvate transaminase

The method for preparing an optically active (R)-amino compound characterized by the method comprising stereoselectively carrying out amino group transfer by action of an (R)-form-specific transaminase in the co-presence of a ketone compound (amino acceptor), and an amino compound (amino donor) of a racemic form or an (R)-form, to give an optically active (R)-amino compound. According to the present invention, it is made possible to easily prepare at a high yield the optically active (R)-amino compounds and the like having an aryl group and the like at their 1-position, which have been conventionally difficult to prepare.

(R)-transaminase from arthrobacter

Process for producing optically active amino compounds

A DNA fragment coding for a decarbamylase protein improved in thermostability as the result of replacement of at least one base of a DNA fragment coding for a decarbamylase protein derived from a microorganism with another base and the resultant replacement of at least one of the corresponding amino acids, and its production process; a vector containing the DNA fragment; a transformant obtained by transformation with the vector; as well as a decarbamylase improved in thermostability and its production process. Also disclosed is a process for producing a D-.alpha.-amino acid, which comprises converting an N-carbamoyl-D-.alpha.-amino acid into the corresponding D-.alpha.-amino acid in an aqueous medium by the action of a decarbamylase having a thermostable temperature of 65.degree. C. or higher; and collecting the D-.alpha.-amino acid produced.

DNA coding for decarbamylase improved in thermostability and use thereof

Process for producing D-amino acids with composite immobilized enzyme

Process for the production of D-.alpha.-amino acids by hydrolysis of the

In a process for the production of a D-.alpha.-amino acid, in which an N-carbamyl-D-.alpha.-amino acid corresponding to the general formula: ##STR1## wherein R represents phenyl, hydroxy-substituted phenyl, substituted or unsubstituted alkyl, or thienyl, is converted by a microbial enzyme in an aqueous medium to a D-.alpha.-amino acid corresponding to the general formula: ##STR2## wherein R is the same as defined above, decarbamylase produced by a microorganism of the genus Comamonas, Blastobacter, Alcaligenes, Sporosarcina, Rhizobium, Bradyrhizobium or Arthrobacter is used as the enzyme converting the N-carbamyl-D-.alpha.-amino acid to the D-.alpha.-amino acid. The conversion of the N-carbamyl-D-.alpha.-amino acids to the D-.alpha.-amino acids is carried out in a neutral to alkaline pH range.

Decarbamylase isolated from comamonas or blastobacter

Decarbamylases are provided capable of producing D-.alpha.-amino acids by hydrolysis of N-carbamyl-D-.alpha.-amino acids. A source of the decarbamylases is recombinant microorganisms produced by gene manipulation methods. Decarbamylases having improved thermostability can be obtained in which amino acids at a thermostability-related site of a natural decarbamylase have been replaced with other amino acids by mutating a DNA fragment encoding the natural decarbamylase. Recombinant DNA is obtained from a vector DNA and a DNA fragment encoding a natural decarbamylase where the nucleic acid sequence encoding an amino acid at a thermostability-related site is replaced with a nucleic acid sequence encoding another amino acid. The recombinant DNA is used to produce transformants that produce thermostable decarbamylases. The decarbamylases may be immobilized in purified, partially-purified or crude form or in the form of decarbamylase-containing microbial cells on a support such as a polymer having ion exchange groups or covalent bonding groups. The presence of an antioxidant such as dithiothreitol results in an immobilized decarbamylase preparation that can be repeatedly used 30 times or more.

Recombinant decarbamylases for producing D-.alpha.-amino acids

The present invention is directed to a gene which is related to a D-N-carbamoyl-.alpha.-amino acid amidohydrolase which is an enzyme capable of converting D-N-carbamoyl-.alpha.-amino acids into D-.alpha.-amino acids; a recombinant plasmid in which a DNA fragment containing the gene is incorporated into a vector; a microorganism belonging to the genus Escherichia, Pseudomonas, Flavobacterium, Bacillus, Serratia, Corynebacterium, or Brevibacterium, which is transformed by incorporating the recombinant plasmid thereinto; a process for the production of D-N-carbamoyl-.alpha.-amino acid amidohydrolases, comprising the steps of cultivating the transformed microorganism and collecting the desired product therefrom; a D-N-carbamoyl-.alpha.-amino acid amidohydrolase obtained by the method; and a process for the production of D-.alpha.-amino acids with the aid of an action of the enzyme. The D-N-carbamoyl-.alpha.-amino acid amidohydrolase can be fixed on a support for immobilization and used as an immobilized enzyme.

Process for production of D-.alpha.-amino acids

A chromosomal DNA sequence which codes for human lymphotoxin, a lymphotoxin expression vector which contains a DNA sequence wherein a chromosomal DNA sequence coding for human lymphotoxin and promoter region which functions in animal cell are linked together, lymphotoxin resistant cell line, transformed animal cell culture which is formed by transforming cultured animal cell with a lymphotoxin expression vector which contains a chromosomal DNA sequence coding for human lymphotoxin and, a process for preparing human lymphotoxin, which comprises transforming cultured animal cell with a lymphotoxin expression vector which contains a chromosomal DNA sequence coding for human lymphotoxin, culturing the transformed cell culture to produce human lymphotoxin, and collecting the human lymphotoxin. According to the present invention, LT which is expected for application as the antitumor agent can be produced effectively in a large amount.

Lymphotoxin DNA, lymphotoxin expression vector

Growing community of inventors

Akashi, Japan

Yasuhiro Ikenaka